Figure 5

Transcriptional regulation by NR5A1 of theca cell marker genes during differentiation in culture. Bovine theca cells were cultivated for 11 days in the presence of minimal amounts of insulin (100 ng/ml) with or without the addition of 10 ng/ml bovine luteinizing hormone (LH) and subsequently infected with adenoviral vectors expressing bovine or murine wild-type NR5A1 or mutants lacking one of the two activating functions (ΔAF-2 or S203A, control: AdEasy adenoviral vector without insert). 72 hours after infection, CYP11A1, STAR, and INSL3 transcripts were quantified by real time-PCR as described in Materials and Methods. The transcript levels in the NR5A1-infected cells are presented in relation to the levels in control-infected cells under identical culture conditions (with or without addition of LH). Thus the bars represent the transcriptional effects of over-expression of NR5A1 wild-type or mutants independent of the stimulatory or inhibitory effects of LH. The results of one representative out of three experiments are shown. Data are expressed as mean ± standard deviation. The groups designated by the letters above the bars differ significantly at the level of P < 0.05.